Homing endonuclease target site specificity defined by sequential enrichment and next-generation sequencing of highly complex target site libraries.
Homing endonuclease target site specificity defined by sequential enrichment and next-generation sequencing of highly complex target site libraries.
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通过高度复杂的靶位点文库的顺序富集和下一代测序定义归巢核酸内切酶靶位点特异性。
DOI：
10.1007/978-1-62703-968-0_12
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
MonnatJr,RaymondJ
中科院分区：
文献类型：
--
作者：
Li,Hui;MonnatJr,RaymondJ
归巢内切酶（HEs）是DNA序列特异性酶，可识别和切割长靶位点（14 - 40bp）以产生双链断裂（DSBs）。它们的高位点识别特异性和结合和裂解的紧密耦合使HEs成为靶向基因组操作的有吸引力的试剂。为了描述HEs的靶点特异性和促进HE工程，我们开发了一种利用部分随机靶点文库和高通量DNA测序对HEs切割特异性进行全面靶点分析的方法。
Homing endonucleases (HEs) are DNA sequence-specific enzymes that recognize and cleave long target sites (14–40 bp) to generate double-strand breaks (DSBs). Their high site recognition specificity and tight coupling of binding and cleavage make HEs attractive reagents for targeted genome manipulation. In order to delineate the target site specificity of HEs and facilitate HE engineering, we have developed a method for comprehensive target site profiling of HEs cleavage specificity using partially randomized target site libraries and high-throughput DNA sequencing.